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March 26, 2014PLoS ONEOpen Access

A Novel Artificial MicroRNA Expressing AAV Vector for Phospholamban Silencing in Cardiomyocytes Improves Ca2+ Uptake into the Sarcoplasmic Reticulum

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Why the study?

Does scAAV6-amiR155-PLBr improve Ca2+ uptake into the sarcoplasmic reticulum with a better safety profile compared to scAAV6-shPLBr in cultured neonatal rat cardiomyocytes?

Population

Cultured neonatal rat cardiomyocytes (CM) isolated from ventricular tissue of 1-3-day-old Wistar rat pups

Comparison

scAAV6-amiR155-PLBr vs scAAV6-shPLBr and control AAV vectors

Design

Preclinical

Follow-up

14 days

Key result

scAAV6-amiR155-PLBr enhanced the Ca2+ transport function of the cardiac SR PLB/SERCA2a system as efficiently as scAAV6-shPLBr while offering a superior safety profile by avoiding proinflammatory protein induction.

Authors

TGTobias GrößlEHElke HammerSBSandra Bien-Möller

Discussion

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Overview

Supports safer microRNA AAV vectors for heart failure calcium handling; leaves open translation to in vivo and human studies.

Structured PICO

Does scAAV6-amiR155-PLBr improve Ca2+ uptake into the sarcoplasmic reticulum with a better safety profile compared to scAAV6-shPLBr in cultured neonatal rat cardiomyocytes?

P
Population
Cultured neonatal rat cardiomyocytes (CM) isolated from ventricular tissue of 1-3-day-old Wistar rat pups
I
Intervention
scAAV6-amiR155-PLBr (AAV vector expressing artificial microRNA directed against PLB under control of a heart-specific hybrid promoter)
C
Comparator
scAAV6-shPLBr (AAV vector expressing short hairpin RNA directed against PLB from an ubiquitously active U6 promoter) and control AAV vectors
O
Outcome
PLB silencing efficiency, SERCA2a-catalyzed Ca2+ uptake into sarcoplasmic reticulum (SR) vesicles, and safety profile (proinflammatory proteins STAT3, STAT1, phosphorylated STAT1, and microRNA expression alterations)surrogate

Main Result

Absolute Event Rate: 77% vs 85%

An artificial microRNA-based AAV vector targeting phospholamban provides comparable improvements in cardiomyocyte calcium handling as shRNA vectors but avoids the induction of proinflammatory STAT proteins, offering a safer gene therapy approach for heart failure.

Limitations

  • In vitro study design limits direct translation to in vivo efficacy and safety

Cite This Study

Größl et al. (2014) studied Heart failure (in vitro model). scAAV6-amiR155-PLBr vs. scAAV6-shPLBr and control vectors was evaluated on Normalized Ca2+ transport activity at submicromolar free Ca2+ concentration (% of maximal value). scAAV6-amiR155-PLBr enhanced the Ca2+ transport function of the cardiac SR PLB/SERCA2a system as efficiently as scAAV6-shPLBr while offering a superior safety profile by avoiding proinflammatory protein induction.

synapsesocial.com/papers/6a178acc8d470cd9925361afhttps://doi.org/10.1371/journal.pone.0092188
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